Hepatitis B X Antigen and P53 Are Associated in Vitro and in Liver Tissues from Patients with Primary Hepatocellular Carcinoma

To test the hypothesis that hepatitis B x antigen (HBxAg) binds to the tumor-suppressor protein p53, immunoprecipitation was carried out with monoclonal anti-x or monoclonal anti-p53 using radiolabeled HBxAg and p53 made by in vitro translation. The results showed that anti-p53 specifically immunoprecipitates HBxAg only in the presence of p53 and that anti-x specifically immunoprecipitates p53 only in the presence of HBxAg. to determine whether HBxAg binds p53 in vivo, immunoprecipitation and Western blot analysis of liver samples from 10 hepatitis B virus (HBV)-infected patients with primary hepatocellular carcinoma (PHC) were carried out. A protein band at 53,000 daltons that specifically immunoprecipitated with a monoclonal anti-x was identified as p53 by Western blotting with a monoclonal anti-p53. Anti-p53 specifically immunoprecipitated bands of 28,000, 17,000 and 13,000 daltons, which were identified as HBxAg polypeptides by Western blotting with anti-HBx. These findings suggest that HBxAg binds to p53 and that this association is important to the development of PHC.